Three-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining

Abstract Tuberculosis (TB) preclinical testing relies on in vivo models including the mouse aerosol challenge model. The only method of determining colony morphometrics of TB infection in a tissue in situ is two-dimensional (2D) histopathology. 2D measurements consider heterogeneity within a single...

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Autores principales:	Robert J. Francis, Gillian Robb, Lee McCann, Bhagwati Khatri, James Keeble, Belinda Dagg, Brad Amos, Francisco J. Salguero, Mei Mei Ho, Anwen Bullen, Gail McConnell, Kirsty MacLellan-Gibson
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c753bc9f75174c4195e6c28918b47a23
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spelling	oai:doaj.org-article:c753bc9f75174c4195e6c28918b47a232021-12-02T11:43:35ZThree-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining10.1038/s41598-020-78640-42045-2322https://doaj.org/article/c753bc9f75174c4195e6c28918b47a232020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78640-4https://doaj.org/toc/2045-2322Abstract Tuberculosis (TB) preclinical testing relies on in vivo models including the mouse aerosol challenge model. The only method of determining colony morphometrics of TB infection in a tissue in situ is two-dimensional (2D) histopathology. 2D measurements consider heterogeneity within a single observable section but not above and below, which could contain critical information. Here we describe a novel approach, using optical clearing and a novel staining procedure with confocal microscopy and mesoscopy, for three-dimensional (3D) measurement of TB infection within lesions at sub-cellular resolution over a large field of view. We show TB morphometrics can be determined within lesion pathology, and differences in infection with different strains of Mycobacterium tuberculosis. Mesoscopy combined with the novel CUBIC Acid-Fast (CAF) staining procedure enables a quantitative approach to measure TB infection and allows 3D analysis of infection, providing a framework which could be used in the analysis of TB infection in situ.Robert J. FrancisGillian RobbLee McCannBhagwati KhatriJames KeebleBelinda DaggBrad AmosFrancisco J. SalgueroMei Mei HoAnwen BullenGail McConnellKirsty MacLellan-GibsonNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-11 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Robert J. Francis Gillian Robb Lee McCann Bhagwati Khatri James Keeble Belinda Dagg Brad Amos Francisco J. Salguero Mei Mei Ho Anwen Bullen Gail McConnell Kirsty MacLellan-Gibson Three-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining
description	Abstract Tuberculosis (TB) preclinical testing relies on in vivo models including the mouse aerosol challenge model. The only method of determining colony morphometrics of TB infection in a tissue in situ is two-dimensional (2D) histopathology. 2D measurements consider heterogeneity within a single observable section but not above and below, which could contain critical information. Here we describe a novel approach, using optical clearing and a novel staining procedure with confocal microscopy and mesoscopy, for three-dimensional (3D) measurement of TB infection within lesions at sub-cellular resolution over a large field of view. We show TB morphometrics can be determined within lesion pathology, and differences in infection with different strains of Mycobacterium tuberculosis. Mesoscopy combined with the novel CUBIC Acid-Fast (CAF) staining procedure enables a quantitative approach to measure TB infection and allows 3D analysis of infection, providing a framework which could be used in the analysis of TB infection in situ.
format	article
author	Robert J. Francis Gillian Robb Lee McCann Bhagwati Khatri James Keeble Belinda Dagg Brad Amos Francisco J. Salguero Mei Mei Ho Anwen Bullen Gail McConnell Kirsty MacLellan-Gibson
author_facet	Robert J. Francis Gillian Robb Lee McCann Bhagwati Khatri James Keeble Belinda Dagg Brad Amos Francisco J. Salguero Mei Mei Ho Anwen Bullen Gail McConnell Kirsty MacLellan-Gibson
author_sort	Robert J. Francis
title	Three-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining
title_short	Three-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining
title_full	Three-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining
title_fullStr	Three-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining
title_full_unstemmed	Three-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining
title_sort	three-dimensional in situ morphometrics of mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/c753bc9f75174c4195e6c28918b47a23
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Three-dimensional in situ morphometrics of Mycobacterium tuberculosis infection within lesions by optical mesoscopy and novel acid-fast staining

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